T5H_TAXCU
ID T5H_TAXCU Reviewed; 499 AA.
AC Q6WG30; Q5XQ40;
DT 28-JUL-2009, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2005, sequence version 2.
DT 03-AUG-2022, entry version 77.
DE RecName: Full=Taxadiene 5-alpha hydroxylase;
DE EC=1.14.14.176 {ECO:0000269|PubMed:15123267};
OS Taxus cuspidata (Japanese yew).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Pinopsida; Pinidae; Conifers II; Cupressales; Taxaceae;
OC Taxus.
OX NCBI_TaxID=99806;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=15123267; DOI=10.1016/j.chembiol.2004.02.022;
RA Jennewein S., Long R.M., Williams R.M., Croteau R.;
RT "Cytochrome p450 taxadiene 5alpha-hydroxylase, a mechanistically unusual
RT monooxygenase catalyzing the first oxygenation step of taxol
RT biosynthesis.";
RL Chem. Biol. 11:379-387(2004).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA].
RA Wang W., Tu J., Cheng K.-D.;
RT "Molecular cloning and sequencing of taxadiene 5-alpha hydroxylase involved
RT in taxol biosynthesis in Taxus chinensis.";
RL Submitted (SEP-2004) to the EMBL/GenBank/DDBJ databases.
CC -!- FUNCTION: Catalyzes the first oxygenation step of taxol biosynthesis.
CC Can use both taxa-4(5),11(12)-diene and taxa-4(20),11(12)-diene as
CC substrate. {ECO:0000269|PubMed:15123267}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=O2 + reduced [NADPH--hemoprotein reductase] + taxa-4,11-diene
CC = H(+) + H2O + oxidized [NADPH--hemoprotein reductase] + taxa-
CC 4(20),11-dien-5alpha-ol; Xref=Rhea:RHEA:14049, Rhea:RHEA-COMP:11964,
CC Rhea:RHEA-COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:30037, ChEBI:CHEBI:30038,
CC ChEBI:CHEBI:57618, ChEBI:CHEBI:58210; EC=1.14.14.176;
CC Evidence={ECO:0000269|PubMed:15123267};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413; Evidence={ECO:0000250};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=48 uM for taxa-4(5),11(12)-diene (for the native enzyme)
CC {ECO:0000269|PubMed:15123267};
CC KM=24 uM for taxa-4(5),11(12)-diene (for the recombinant enzyme)
CC {ECO:0000269|PubMed:15123267};
CC KM=27 uM for taxa-4(20),11(12)-diene (for the native enzyme)
CC {ECO:0000269|PubMed:15123267};
CC KM=16 uM for taxa-4(20),11(12)-diene (for the recombinant enzyme)
CC {ECO:0000269|PubMed:15123267};
CC -!- PATHWAY: Alkaloid biosynthesis; taxol biosynthesis; taxa-4(20),11-dien-
CC 5alpha-ol from geranylgeranyl diphosphate: step 2/2.
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000305}; Single-pass membrane
CC protein {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAU93341.1; Type=Frameshift; Evidence={ECO:0000305};
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DR EMBL; AY289209; AAQ56240.2; -; mRNA.
DR EMBL; AY741375; AAU93341.1; ALT_FRAME; mRNA.
DR AlphaFoldDB; Q6WG30; -.
DR SMR; Q6WG30; -.
DR KEGG; ag:AAQ56240; -.
DR BioCyc; MetaCyc:MON-13393; -.
DR BRENDA; 1.14.14.176; 6225.
DR SABIO-RK; Q6WG30; -.
DR UniPathway; UPA00842; UER00807.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0050604; F:taxadiene 5-alpha-hydroxylase activity; IEA:UniProtKB-EC.
DR GO; GO:0042617; P:paclitaxel biosynthetic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 1.10.630.10; -; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR002401; Cyt_P450_E_grp-I.
DR InterPro; IPR036396; Cyt_P450_sf.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00463; EP450I.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; SSF48264; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW Heme; Iron; Membrane; Metal-binding; Monooxygenase; Oxidoreductase;
KW Signal-anchor; Taxol biosynthesis; Transmembrane; Transmembrane helix.
FT CHAIN 1..499
FT /note="Taxadiene 5-alpha hydroxylase"
FT /id="PRO_0000380688"
FT TRANSMEM 22..42
FT /note="Helical; Signal-anchor"
FT /evidence="ECO:0000255"
FT BINDING 445
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="axial binding residue"
FT /evidence="ECO:0000250"
FT CONFLICT 31
FT /note="A -> S (in Ref. 2; AAU93341)"
FT /evidence="ECO:0000305"
FT CONFLICT 175
FT /note="S -> N (in Ref. 2; AAU93341)"
FT /evidence="ECO:0000305"
FT CONFLICT 249
FT /note="K -> T (in Ref. 2; AAU93341)"
FT /evidence="ECO:0000305"
FT CONFLICT 413
FT /note="N -> S (in Ref. 2; AAU93341)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 499 AA; 56558 MW; D1503DA678CA4619 CRC64;
MDALYKSTVA KFNEVTQLDC STESFSIALS AIAGILLLLL LFRSKRHSSL KLPPGKLGIP
FIGESFIFLR ALRSNSLEQF FDERVKKFGL VFKTSLIGHP TVVLCGPAGN RLILSNEEKL
VQMSWPAQFM KLMGENSVAT RRGEDHIVMR SALAGFFGPG ALQSYIGKMN TEIQSHINEK
WKGKDEVNVL PLVRELVFNI SAILFFNIYD KQEQDRLHKL LETILVGSFA LPIDLPGFGF
HRALQGRAKL NKIMLSLIKK RKEDLQSGSA TATQDLLSVL LTFRDDKGTP LTNDEILDNF
SSLLHASYDT TTSPMALIFK LLSSNPECYQ KVVQEQLEIL SNKEEGEEIT WKDLKAMKYT
WQVAQETLRM FPPVFGTFRK AITDIQYDGY TIPKGWKLLW TTYSTHPKDL YFNEPEKFMP
SRFDQEGKHV APYTFLPFGG GQRSCVGWEF SKMEILLFVH HFVKTFSSYT PVDPDEKISG
DPLPPLPSKG FSIKLFPRP
